KnockoutTM SR无血清培养基支持小鼠精原干细胞的短期存活(简报)

精原干细胞(spermatogonial stem cells,SSCs)是睾丸内具有自我复制和分化为精子潜能的干细胞,它的体外培养是精子发生机理研究和制作转基因动物等的新途径[1,2].近几年的研究表明,SSCs在体外的自我增殖需要GDNF(glial cell line-derived neu-rotrophie factor)因子和饲养层细胞等的支持[3-10].并且睾丸支持细胞(Sertoli's cells)和血清都导致培养的SSCs分化[1,6].因此,使用无血清培养基培养高度纯化的SSCs是培养成败的关键之一.

SERUM-FREE KnockouTM SR MEDIUM SUPPORTS THE SHORT-TIME VIABILITY OF MOUSE SPERMATOGONIAL STEM CELLS

Spermatogonial stem cells (SSCs), the only stem cells that are capable of transmitting genetic information to subsequent generations in adult animals,have abilities to self-renew and differentiate into spermatozoa. Therefore, SSCs are not only the study object of stem cell biology, but also the valuable resource of in vitro spermatogenesis, gene analysis and functional genomics. In the present study, we selected the mouse SSCs from mice on STO (SIM mouse embryo-derived thioguanine and ouabain resistant) feeders by differential adherence selection in DMEM/F12 containing 5% FBS, and used a serum-free defined medium prepared with Knockout SR basal medium to culture SSCs. Our results showed that enriched SSCs could be maintained for a short of time and form colonies, but the proliferation of SSCs was unconspicuous, suggesting that some factors that are detrimental to the self-renewal of SSCs possibly existed in the Knockout SR basal medium. However, the use of KSR medium, which was widely used in the culture of embryonic stem cells (ESCs), in the SSC maintenance was the first time, and our results indicated that the Knockout SR basal medium don't appropriate for the long-time culture of SSCs.